Objective To establish a duplex droplet digital polymerase chain reaction (ddPCR) detection method for Japanese encephalitis virus (JEV) and West Nile virus (WNV).Methods Based on the designed primers and probes of JEV and WNV, a duplex ddPCR detection system for JEV and WNV was established. Its sensitivity, specificity, and repeatability were explored. The sensitivity was compared with the number of cycles required for the fluorescent signal to cross the threshold in each reaction tube of dual quantitative PCR.Results The detection sensitivity of the duplex ddPCR detection system could reach 102 copies/μl for both JEV and WNV, with good specificity and repeatability. No cross-reactivity was observed with the Dengue virus, Chikungunya virus, Zika virus, Tick-borne encephalitis virus, and human genome.Conclusion The established duplex ddPCR method shows high sensitivity and specificity for JEV and WNV detection, which provides a solution for detection for the two viruses in different scenarios.
ZHANG Jun-feng, ZHANG Ya-li, WANG Rui-chen, LU Yang, ZHANG Tian-zi, FU Shi-hong, YIN Qi-kai, LI Fan, HE Ying, NIE Kai, MA Chao-feng, LIANG Guo-dong, HU Rui-ping, XU Song-tao, WANG Huan-yu
. Establishment of a duplex droplet digital PCR assay for Japanese encephalitis and West Nile viruses[J]. Chinese Journal of Vector Biology and Control, 2023
, 34(3)
: 285
-290
.
DOI: 10.11853/j.issn.1003.8280.2023.03.001
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